The objective of this study was to apply high resolution melting
analysis in the detection of complex vertebral malformation (CVM)
carriers in Hosltein cattle. A total of 47 animals of Holstein cattle were
included in this study. Genomic DNA was extracted from blood, hair
follicles and sperm by commercial QIAamp® DNA Mini kit. The
amplification and high resolution melting analysis (HRMA) was done by
commercial SensiMixTM HRM kit. The confirmation of sensitivity of this
method was done by PCR-PIRA method and sequencing. Four samples
of heterozygous genotype GT for causal mutation in the bovine solute
carrier family 35 member 3 gene (SLC35A3), which is responsible for
CVM disease, were detected. Our results demostrated that the use of
HRMA for genotyping of mutant allele T for SLC35A3 gene in Holstein
cattle is an effective method for the selection of carriers of CVM
disease.